Using nutritionally deficient mutants of a series of gram-negative coccobacilli, including species of Moraxella and Neisseria, transformation assay systems will be devised to permit definitive identification of a variety of bacteria belonging to different genetically homogeneous groups. These assays will be developed for routine diagnostic use in hospital bacteriology laboratories, and also in research laboratories. In order to improve the specificity of the transformation assay system already established for Neisseria gonorrhoeae, mutants will be sought in genes that are not common to non-gonorrhoeae neisserias. A transformation assay will also be devised for non-gonorrhoea neisserias. Studies of the nature of colony type variation in Neisseria gonorrhoeae will be undertaken to determine whether or not plasmid DNA, interacting with chromosomal DNA, plays a role in this unusual variation. Efforts will be made to determine the simplest nutritional requirements for some neisserias. The biochemical nature of the reaction that makes it necessary for neisserias to be incubated in an atmosphere having a high CO2 concentration for initiation of growth will be investigated. Certain aspects of the mechanisms of competency for transformation will be studied. Specifically, attempts will be made to mutate or transduce non-competent strains to competency. Also to be studied will be the mechanisms whereby competent strains become non-competent spontaneously. Using transformation, the possibility of constructing extensively hybridized strains containing chromosomal DNA from both Acinetobacter and Moraxella osloensis will be explored. Current studies of metabolic pathways for gluconeogenesis in Acinetobacter will be extended making use of mutants defective in relevant enzyme activities in order to determine the physiological significance of the pathways under investigation. This work, on the mechanisms of gluconeogenesis, will be extended to other gram-negative coccobacilli, particularly to those that are unable to utilize sugars as carbon sources for growth.